Hydroxylated 1 alpha, 2 alpha-methylene steroids

ABSTRACT

Steroids of the formula ##STR1## wherein R 1  and R 2  are identical or different and each is hydrogen or the residue of an organic or inorganic acid, 
     and for the esters (R 1  or R 2  is not H) capable of salt formation, the salts thereof, possess valuable pharmacological properties, e.g., good antiandrogenic activity with low progestational activity.

BACKGROUND OF THE INVENTION

The present invention relates to pharmaceutically active 1α,2α-methylenesteroids.

SUMMARY OF THE INVENTION

It is an object of this invention to provide new steroids possessinghigh antiandrogenic activity with minor progestational activity.

Upon further study of the specification and appended claims, furtherobjects and advantages of this invention will become apparent to thoseskilled in the art.

These objects have been attained by providing hydroxylated1α,2α-methylene steroids of Formula I ##STR2## wherein R₁ and R₂ areidentical or different and each is hydrogen or the residue of an organicor inorganic acid, and for the esters (R₁ and/or R₂ is not H) capable ofsalt formation, the salts thereof.

DETAILED DISCUSSION

To form the esters of Formula I, organic acids customarily used insteroid chemistry can be employed. Such acids include, for example,organic carboxylic and sulfonic acids of 1-18, preferably 1-12 carbonatoms. Such acids include those of the aliphatic, cycloaliphatic,aromatic, aromatic-aliphatic, or heterocyclic series, (S, O or N heteroatoms), any of which can be unsaturated and/or polybasic and/orconventionally substituted. Examples of such substituents include alkyl,hydroxy, alkoxy, oxo, amino, or halogen (F, Cl, Br, I). Althoughalkanoic and alkanesulfonic acids of 1-12 carbon atoms are especiallypreferred, contemplated equivalents are acids of all other carboxylicand sulfonic acids, e.g., short-chain hydroxycarboxylic acids of 2-6carbon atoms esterified with further carboxylic or sulfonic acids of2-18 carbon atoms, e.g., glycolic acid O-nonanoate, glycolic acidO-trideconoate, glycolic acid O-hexadecanoate etc. Contemplated asequivalents of the carboxylic or sulfonic acids are also those ofcarbocyclic aryl or alkaryl acids, e.g., containing 7-18 carbon atomsand preferably 1 or 2 rings; aralkyl acids, e.g., containing 8-18 carbonatoms, heterocyclic acids of 1 ring, e.g., containing 4-9 carbon atomsand 1-2 hetero atoms (N, O, S).

Examples of suitable carboxylic acids include: formic acid, acetic acid,propionic acid, butyric acid, isobutyric acid, valeric acid, isovalericacid, caproic acid, enanthic acid, caprylic acid, pelargonic acid,capric acid, undecylic acid, trimethylacetic acid, diethylacetic acid,cyclopentylpropionic acid, cyclohexylacetic acid, phenylacetic acid,benzoic acid, glycolic acid, glycolic acid O-undecanoate, mono-, di-,and trichloroacetic acid, oxalic acid, malonic acid, succinic acid,adipic acid, pimelic acid, suberic acid, aminoacetic acid,diethylaminoacetic acid, diethylaminopropionic acid, piperidinoaceticacid, morpholinoacetic acid, nicotinic acid, isonicotinic acid,furan-2-carboxylic acid, and similar acids. Compounds of general FormulaI also primarily include the physiologically compatible salts of thedicarboxylic acid monoesters with bases and the physiologicallycompatible addition salts of the aminocarboxylic acid esters with acids.

Examples of sulfonic acids suitable for ester formation include:methanesulfonic acid, ethanesulfonic acid, isopropanesulfonic acid,butanesulfonic acid, cyclopropanesulfonic acid, cyclopentanesulfonicacid, benzenesulfonic acid, p-toluenesulfonic acid,p-chlorobenzenesulfonic acid, N,N-dimethylaminosulfonic acid,N,N-diethylaminosulfonic acid, pyrrolidino-, piperidino-, piperazino-,N-methylpiperazino-, morpholinosulfonic acids, and the like. From theaminosulfonic acid esters, the physiologically compatible water-solubleacid addition salts can be produced.

Also suitable, particularly in the 15-position, are the esters ofinorganic acids such as the hydrohalic (Cl, Br, I) acids, preferablysulfuric and phosphoric acids, as well as the physiologically compatiblesalts of the hemisulfate and monophosphoric acid esters with bases.

Physiologically compatible salts of the aforementioned bases includetheir alkali- and alkaline earth-metals, especially sodium salts. Forthe formation of the aforementioned acid addition salts, suitablephysiologically compatible salts are those acids which areconventionally employed in connection with organic nitrogen compounds.Suitable such acids for salt formation include, for example,hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid,propionic acid, lactic acid, tartaric acid, succinic acid, citric acid,benzoic acid, salicylic acid, nicotinic acid, adamantanecarboxylic acid,etc.

A process for the preparation of the hydroxylated 1α,2α-methylenesteroids of Formula I comprises fermenting compounds of Formula II##STR3## wherein R₁ is as defined above, with a bacterial culture of thegenus Bacillus or with a fungal culture of the genus Mucor, and,esterifying any free hydroxy groups in the 17- and/or 15-positions whenrequired to produce the desired R₁ and R₂ moieties.

When R₁ =H, Formula II represents the known steroid, cyproterone. Thestarting material cyproterone esters of Formula II can be prepared inaccordance with the esterification procedures discussed below, ifdesired, or the corresponding esterification can be performed after the15β-hydroxylation.

The 15β-hydroxylation is accomplished using a bacterial culture of thegenus Bacillus or with a fungal culture of the genus Mucor. Suitable forthis fermentation are the bacterial strain, for example, Bacillusmegaterium (ATCC 13 368) and the fungal strain, for example, Mucorgriseocyanus (ATCC 1207 b).

The fermentation is conducted under the same conditions employed inconventional fermentative conversions of steroids using bacterial andfungal cultures. For the fermentative conversions see W. Charney and H.L. Herzog "Microbial Transformation of Steroids", Academic Press, NewYork and London, 1967.

Thus, initially, the most favorable fermentation conditions aredetermined by conventional routine preliminary tests such as, forexample, the selection of the most favorable nutrient medium; thesuitable substrate solvent; the substrate concentration; the technicalconditions, such as temperature, aeration conditions, pH value, etc.;and the optimum periods of germination, addition of substrate andsubstrate contact with the enzyme of the microorganism. These latterdeterminations are done analytically, especially by thin-layerchromatography.

It has been found, in this connection, that it is advantageous toutilize concentrations of about 50-1,000 mg of substrate per liter ofnutrient medium. The pH value is preferably in the range of 5-7. Theculturing temperature is in the range of 20°-40° C., preferably 25°-35°C., and the pressure 0-1 atm. gauge. For aeration, about 1 liter of airper minute per liter of culture broth is added. The conversion of thesubstrate is suitably observed by analyzing sample extracts usingthin-layer chromatography. IN general, satisfactory quantities ofhydroxylated steroid have formed after 50-100 hours.

After the fermentation is complete, the fermentation products areisolated conventionally. The isolation can be done, for example, byextracting the fermentation batches with a polar solvent which is notwater-soluble, such as ethyl acetate, butyl acetate, or methyl isobutylketone; concentrating the extracts; and purifying the thus-obtainedcrude products optionally by chromatography and/or crystallization.

For the esterification step, conventional steroid chemistry processescan be utilized as disclosed for example in U.S. Pat. No. 4,011,314,whose disclosure is incorporated by reference herein. Since thesecondary hydroxy group in the 15-position and the tertiary hydroxygroup in the 17-position exhibit different reactivities, the hydroxygroups can also be esterified in stages. Moreover, it is frequentlyuseful to start with compounds already esterified in the 17α-position,particularly if relatively drastic conditions are required for theesterification of the 15-hydroxy group, such as, for example, in theesterification with inorganic acids or when producing the aminoacylatesfrom the corresponding chloroacylates. The partial esterification in the15-position is preferably conducted in the presence of an alkalinecatalyst at room temperature. Suitable catalytic bases are preferablytertiary amines, e.g. triethylamine, pyridine, collidine, optionallywith the addition of 4-dimethylaminopyridine. Suitable esterifyingagents are reactive acid derivatives, such as acid halogenides oranhydrides of monocarboxylic acids.

The esterification of the hydroxy group in the 17-position with amonocaroxylic acid may be conducted by utilizing fully conventionalmethods, see for example U.S. Pat. No. 3,749,742 and U.S. Pat. No.2,753,360. One example is the reaction with the anhydride or halogenideof the desired acid in the presence of a strongly acidic catalyst, suchas perchloric acid, p-toluenesulfonic acid, trifluoroacetic acid, atroom temperature or in the presence of an alkaline catalyst, such aspyridine, collidine, quinoline, opitionally with the addition of4-dimethylaminopyridine, at temperatures of from above room temperatureto the boiling point of the base.

The esterification of the hydroxy groups in the 15- and 17-positions canalso be effected with a free carboxylic acid in the presence oftrifluoroacetic anhydride, see U.S. Pat. No. 3,383,394.

If the 3-enol ester is formed simultaneously during the acidicesterificatiion, the enol ester grouping can be selectively split off ina conventional manner, for example by treatment with mineral acids oralso with paratoluenesulfonic acid in an alcoholic solution, see U.S.Pat. No. 2,753,360.

The esterification of the free hydroxy groups with dicarboxylic acids isconducted, according to a preferred embodiment, with a mixture oftrifluoroacetic anhydride and the desired dicarboxylic acid. Theesterification is advantageously carried out at 10°-50° C. in thepresence of a solvent inert to the reactants. Suitable solvents includewater-miscible solvents, such as tetrahydrofuran or dioxane, as well aswater-immiscible solvents, such as benzene, see U.S. Pat. No. 3,525,755.

The water solubility of suitable salts of the compounds esterified withone or two dicarboxylic acids is of special significance. For purposesof such salt formation, the acidic monoesters are dissolved, forexample, in alcohol and neutralized with the base, especially sodiumhydroxide solution. A compound monoacylated in the 17-position with acarboxylic acid can be converted into the 15-sulfonic acid ester byconventionally reacting the 17-monoacyl compound in the presence of atertiary amine with a sulfonic acid halogenide at room temperature. If a15,17-dihydroxy compound is reacted with a sulfonic acid halogenide inthe presence of a tertiary amine at room temperature, the15,17-disulfonic acid ester is obtained.

For conducting the esterification with an inorganic acid and for thepreparation of aminoacylates, compounds which have already beenesterified in the 17α-position are suitably used as the startingmaterials. Thus, 15-hemisulfate alkali metal salts are prepared from the15-hydroxy compounds according to methods known per se, e.g., asdescribed in British Pat. No. 1,286,093. For example, initially,pyridine-sulfur trioxide adduct can be used to convert the 15-hydroxycompounds into the acidic 15-sulfuric acid esters, and the latter arethen conventionally converted into the physiologically compatible saltswith bases. Suitable bases include alkali metal hydroxides andalcoholates, especially of sodium.

The 15-aminoacylates of this invention, such as, for example, thediethylamino-, piperidino-, and morpholinoacetates, can be obtainedaccording to known methods by reaction of the corresponding 15-hydroxycompounds with a halofatty acid anhydride or halogenide in the presenceof an organic base, such as pyridine, by way of the 15-haloacylate andsubsequent reaction with the desired amine, see British Pat. No.1,286,093. Preferred amines are secondary aliphatic and cyclic amines,such as diethylamine, pyrrolidine, piperidine, piperazine,N-methylpiperazine, morpholine, etc. The reaction of the 15-haloacylatewith the amine takes place simply by heating the components together.Suitably, excess amine serves as the solvent. Essentially, the reactioncan also be conducted in a dilute solution. The thus-obtained15-aminoacylates can be converted into the corresponding acid additionsalts by following conventional methods.

The hydroxylated 1α,2α-methylene steroids of Formula I arepharmacologically active compounds. For example, the novel compoundspossess a strong antiandrogenic activity with minor progestationalactivity.

To determine the antiandrogenic activity, the inhibition of thetestosterone propionate caused increase in the weight of the seminalvesicles and prostates of rats was determined upon subcutaneousapplication. As demonstrated by the following Table 1, a compound ofthis invention,17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione(A) has the same strong antiandrogenic activity as the strongantiandrogen,17-acetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione (B). Theantiandrogenic activity was tested on male castrated rats as describedin Acta endocr. (Kbh.) Suppl. 90 (1964) 139.

                  TABLE 1                                                         ______________________________________                                                        Testosterone Propionate                                              Dose     Inhibition                                                    Compound (mg/day)   Seminal Vesicle                                                                             Prostate                                    ______________________________________                                        A        1.0        72%           72%                                         B        1.0        73%           74%                                         ______________________________________                                    

In the Clauberg test after subcutaneous application to rabbits, the15β-OH-compound A of this invention is ineffective at a dosage of 0.03mg (McPhail=1.1). The conventional antiandrogen B is still effective ata dosage of only 0.003 mg (McPhail=1.5). The gestagenic effect waschecked in the usual Clauberg test (Zbl. Gynak. Vol. 54 (1930) 2757 andJ. Physiol. (London) Vol. 83 (1934) 10 and 145).

                  TABLE 2                                                         ______________________________________                                        Compound    Dose (mg)     McPhail                                             ______________________________________                                        A           0.03          1.1                                                 B           0.003         1.5                                                 ______________________________________                                    

On the basis of such favorably opposed antiandrogenic and progestationalactivities, the compounds of this invention are particularly wellsuitable for the treatment of diseases caused by androgens or dependenton androgens in mammals, including humans. Thus, the compounds can beutilized, for example, in the form of the water-soluble ester salts forthe local treatment of acne and seborrhea.

The pharmacologically active compounds of Formula I can be processed inaccordance with conventional methods of galenic pharmacy to producemedicinal agents. Conventional excipients are pharmaceuticallyacceptable organic or inorganic carrier substances suitable forparenteral, enteral or topical application which do not deleteriouslyreact with the active compounds. Suitable pharmaceutically acceptablecarriers include but are not limited to water, salt solutions, alcohols,vegetable oils, polyethylene glycols, gelatine, lactose, amylose,magnesium stearate, talc, silicic acid, viscous paraffin, perfume oil,fatty acid monoglycerides and diglycerides, pentaerythritol fatty acidesters, hydroxymethylcellulose, polyvinyl pyrrolidone, etc. Thepharmaceutical preparations can be sterilized and if desired mixed withauxiliary agents, e.g., lubricants, preservatives, stabilizers, wettingagents, emulsifiers, salts for influencing osmotic pressure, buffers,coloring, flavoring and/or aromatic substances and the like which do notdeleteriously react with the active compounds.

For parenteral application, particularly suitable are solutions,preferably oily or aqueous solutions, as well as suspensions, emulsions,or implants, including suppositories. Ampoules are convenient unitdosages.

For enteral application, particularly suitable are tablets, dragees, orcapsules having talc and/or a carbohydrate carrier or binder or thelike, the carrier preferably being lactose and/or corn starch and/orpotato starch. A syrup, elixir or the like can be used wherein asweetened vehicle is employed.

For topical application, the compounds of this invention are employed asnon-sprayable forms, viscous to semi-solid or solid forms comprising acarrier indigenous to topical application and having a dynamic viscositypreferably greater than water. Suitable formulations include but are notlimited to solutions, suspensions, emulsions, creams, ointments,powders, linaments, salves, aerosols, etc., which are, if desired,sterilized or mixed with auxiliary agents, e.g., preservatives,stabilizers, wetting agents, buffers, or salts for influencing osmoticpressure, etc. For topical application, also suitable are sprayableaerosol preparations wherein the active ingredient, preferably incombination with a solid or liquid inert carrier material, is packagedin a squeeze bottle or in admixture with a pressurized volatile,normally gaseous propellant, e.g., a freon. Usually, the activecompounds of the invention are incoroporated in topical formulations ina concentration of about 0.001 to 5 weight percent.

Sustained release compositions can be formulated including those whereinthe active compound is protected with differentially degradablecoatings, e.g., by microencapsulation, multiple coatings, etc.

Generally, the compounds of this invention are dispensed in unit dosageform comprising 10-100 mg in a pharmaceutically acceptable carrier perunit dosage.

The dosage of the compounds according to this invention is 10-500 mg/daywhen administered to human patients as an antiandrogenic agent. Suitablemodes of administration are in the same manner as the knownantiandrogenic agent B.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following preferred specific embodiments are,therefore, to be construed as merely illustrative, and not limitative ofthe remainder of the disclosure in any way whatsoever. In the followingexamples, all temperatures are set forth uncorrected in degrees Celsius;unless otherwise indicated, all parts and percentages are by weight.

EXAMPLE 1

A 2-liter Erlenmeyer flask containing 500 ml. of a nutrient solution of0.1% peptone, 0.2% corn steep liquor, 0.5% glucose, and 0.5% yeastextract, sterilized for 30 minutes in an autoclave at 120° C.--adjustedto pH 7.2--is inoculated with a slanted-tube agar culture of Bacillusmegaterium (ATCC 13 368) and shaken for 48 hours at 30° C. on a rotaryshaker.

A 20-liter fermentor filled with 15 liters of a medium of the samecomposition as the shaker flask subculture, sterilized at 121° C. andunder 1.1 atmospheres gauge, is then inoculated with this subculture.With the addition of several cubic centimeters of Silicone SH as anantifroth agent, the culture is germinated at 29° C. under aeration (10l./min.), a pressure of 0.7 atm. gauge, and under agitation (220 r.p.m.)for 24 hours.

Under sterile conditions, 1.8 l. of culture broth is then withdrawn fromthis preliminary fermentor culture, and a 50-liter fermentor containing28 l. of sterilized nutrient medium of the same composition as thepreliminary fermentor culture is inoculated with this culture broth. Thelatter is germinated under the same conditions as in the preliminaryfermentor (29° C., 30 l./min. of air, 0.7 atm. gauge, 220 r.p.m.). Aftera growth time of 12 hours, a solution of 6 g. of cyproterone acetate(17-acetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione) in 45ml. of dimethylformamide is added, and the culture is further agitatedand aerated.

After a contact period of 88 hours, the optimum of conversion has beenreached. The fermentor content is, at this point in time, extractedfirst with one-half and then once more with one-third of the volume withmethyl isobutyl ketone by agitation. The extracts are combined andconcentrated in a forced circulation evaporator at 28° C. under a vacuumto 1 liter. The concentrate is evaporated to dryness in the forcedcirculation evaporator at a bath temperature of 50° C. The residue istaken up in methanol; the undissolved silicone oil (antifroth agent) isseparated, and the methanolic solution is evaporated to dryness undervacuum.

To purify the hydroxylation product and to separate unreacted startingmaterial, the residue is chromatographed over a silica gel column andeluted with the aid of a linear gradient hexane-hexane/acetone. Aftercrystallization from acetone/isopropyl ether, the pure17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dionemelts at 276°-277° C. The yield is 43%.

EXAMPLE 2

Under the culturing and fermentation conditions of Example 1, a 50-literfermentor is prepared with 28 l. of a Bacillus megaterium culture and,after a culture period of 12 hours, combined with 6 g. of cyproterone(6-chloro-17-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione),dissolved in 400 ml. of dimethyl sulfoxide. After a contact period of 60hours, the content of the fermentor is extracted with methyl isobutylketone and worked up as described in Example 1. The thus-obtained crudeproduct is chromatographed over a silica gel column with the use of thesolvent gradient methylene chloride-methylene chloride/acetone andrecrystallized from acetone/hexane, thus obtaining in a 36% yield pure6-chloro-15β,17-dihydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,m.p. 251°-253° C.

EXAMPLE 3

(a) 40 g. of adipic acid is suspended in 300 ml. of benzene, and 120 ml.is distilled off to remove water. The mixture is then cooled to 5° C.and, at this temperature, 75 g. of trifluoroacetic anhydride is addedunder agitation. The reaction mixture is stirred for about 1 hour atroom temperature, thus producing a clear solution.

Thereafter 14 g. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneis added thereto and the solution, which is now discolored to a darkbrown, is agitated for 5 hours at room temperature. To work up thereaction mixture, the solution is stirred into 1 liter of ice water,stirred for 15 minutes under ice cooling, and the pH value is adjustedto pH 5 with 5 N NaOH solution. Subsequently the mixture is extractedthree to five times with respectively 1/2 liter of ethyl acetate; theextracts are combined and concentrated to about 1 liter by evaporationunder vacuum.

To purify the hemiadipate, the ethyl acetate solution is extracted fivetimes with respectively 750 ml. of 3% NaHCO₃ solution, and the combinedNaHCO₃ solutions are washed twice with respectively 300 ml. of ethylacetate. The NaHCO₃ solution is acidified under agitation with 5 N HClto pH 3.5-4.5 and extracted four times with respectively 1/2 liter ofethyl acetate. The combined ethyl acetate phases are finally washed freeof adipic acid with a small amount of distilled water and dried over Na₂SO₄. Upon evaporation under vacuum, the 15-hemiadipate remains in theform of a yellow, oily residue (13.5 g.). After crystallization fromether, the pure17-acetoxy-15β-adipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dionemelts at 172°-174° C.

(b) 10.2 g. of17-acetoxy-15β-adipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dioneis dissolved in 50 ml. of ethanol, the solution is cooled to about 5° C.and adjusted to pH 8.0 at this temperature under agitation with the aidof N/10 NaOH. The mixture is then filled up with distilled water to avolume of 150 ml., 1 g. of activated carbon is added, and the mixture isagitated for 1 hour at room temperature. The mixture is then filteredtwice through a folded filter, and the filtrate is distilled at a bathtemperature of no higher than 30°-35° C. under vacuum until the mainquantity of the alcohol has been removed. The colorless solution is thenfrozen in a refrigerating bath of -70° C. under rotary motion in a roundflask and subjected to freeze-drying. Yield: 9.8 g. of17-acetoxy-15β-adipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,sodium salt, m.p. 180°- 200° C. (under decomposition).

EXAMPLE 4

Under the conditions of Example 3(a), 1.5 g. of malonic acid in benzeneis reacted with 2 ml. of trifluoroacetic anhydride. After the additionof 1 g. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,the mixture is stirred for 15 minutes and then poured into ice water.After adjusting the pH to 5, the mixture is extracted repeatedly withether and the residue of the combined ether solutions, evaporated todryness, is chromatographed over a silica gel column with the use of alinear solvent gradient methylene chloridemethylene chloride/acetone.After concentration of the main fraction,17-acetoxy-6-chloro-15β-malonyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneremains as a light-colored oil (665 mg.) which, after digestion, iscrystallized with isopropyl ether (m.p. 185°-189° C.). The thus-obtainedhemimalonate is converted analogously to Example 3(b) into thecorresponding sodium salt, thus obtaining 600 mg. of17-acetoxy-6-chloro-15β-malonyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,sodium salt, m.p. 193°-195° C.

EXAMPLE 5

Under the conditions of Example 3(a), 4 g. of adipic acid in 30 ml. ofbenzene is reacted with 5 ml. of trifluoroacetic anhydride. Then 3 g. of6-chloro-15β,17-dihydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione isadded thereto and the mixture is agitated for 3 hours at 150° C. Afterthe mixture has been worked up as described in Example 3(a), thethus-obtained crude product is once more chromatographed over a silicagel column (gradient: methylene chloride-methylene chloride/acetone),thus obtaining15β-adipoyloxy-6-chloro-17-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dionein the form of a hemicrystalline foam (2.2 g.) melting within a range of100°-150° C.

The thus-obtained hemiadipate is now converted into the sodium saltanalogously to Example 3(b), so that 2 g. of15β-adipoyloxy-6-chloro-17-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,sodium salt, is obtained, m.p. 160°-180° C. (under decomposition).

EXAMPLE 6

Under the conditions of Example 3(a), 2.6 g. of adipic acid in 20 ml. ofbenzene is reacted with 3.3 ml. of trifluoroacetic anhydride.Thereafter, 2 g. of6-chloro-15β,17-dihydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione isadded thereto, and the mixture is stirred for 5 hours at roomtemperature. After the reaction mixture has been worked up as describedin Example 3(a), the thus-obtained crude product is chromatographed, forpurposes of further purification, once again over a silica gel columnand eluted by means of the solvent gradient methylene chloride-methylenechloride/acetone and with the addition of several drops of concentratedhydrochloric acid. The main fraction is evaporated to dryness and theresidue is crystallized from ether/ethyl acetate/hexane. The pure15β,17-diadipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione(1.3 g.) melts at 130°-140° C. under decomposition.

The thus-obtained dihemiadipate is now converted into the disodium saltanalogously to Example 3(b), so that finally 1.3 g. of15β,17-diadipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,disodium salt, is obtained, melting at 200° C. under decomposition.

EXAMPLE 7

500 mg. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneis dissolved in 20 ml. of pyridine; 10 ml. of acetic anhydride and 500mg. of 4-dimethylaminopyridine are added thereto and the mixture isstirred for 16 hours under nitrogen at room temperature. Then themixture is poured into ice-cooled 8% aqueous sulfuric acid, extractedwith ethyl acetate, washed neutral, and evaporated under vacuum todryness. After crystallizing the residue from ethyl acetate/isopropylether in the presence of activated carbon, the pure15β,17-diacetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dionemelts at 285°-287° C. Yield: 440 mg.

EXAMPLE 8

10 ml. of absolute pyridine is cooled to -15° C. and 0.8 ml. of sulfurtrioxide is gradually added dropwise under agitation so that theinternal temperature does not rise above -5° C. Into this suspension isintroduced 2.5 g. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dionein 5 ml. of pyridine. The reaction mixture is stirred for 30 minutes atroom temperature, then diluted with 50 ml. of water, and finallyadjusted to pH 9 with 9 ml. of 1 N NaOH. To remove the pyridine, themixture is extracted with methylene chloride, the pH is set to 8 with 1N NaOH, and the mixture is evaporated under vacuum. The residue isdissolved in methanol, the sodium sulfate still undissolved in filteredoff, the solution is treated with activated carbon, and, after anotherfiltering step, evaporated once again to dryness, thus obtaining 2.1 g.of sodium(17-acetoxy-6-chloro-1α,2α-methylene-3,20-dioxo-4,6-pregnadien-15β-yl)sulfate, m.p. 88°-90° C.

EXAMPLE 9

A solution of 2.5 g. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dionein 12 ml. of pyridine is stirred at 0°-5° C. under nitrogen and combinedwith 1.5 g. of chloroacetic anhydride in 9 ml. of absolute ether. Themixture is stirred for another 5 hours, and the bath temperature isallowed to rise gradually to room temperature. Then 0.3 ml. of water isadded, and the product is precipitated after 5-10 minutes by pouring thereaction solution into ice water. The product is vacuum-filtered andwashed, in succession, with 5% hydrochloric acid, dilute sodiumbicarbonate solution, and water.

The yield is 2.9 g. of a crude product. After recrystallizing twice fromether, the pure17-acetoxy-6-chloro-15β-chloroacetoxy-1α,2α-methylene-4,6-pregnadiene-3,20-dionemelts at 225°-228° C.

Under nitrogen, 1.5 g. of17-acetoxy-6-chloro-15β-chloroacetoxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneis heated to boiling with 40 ml. of diethylamine for 3 hours. Aftercooling to room temperature, the mixture is diluted with chloroform andevaporated under vacuum. The residue is taken up in chloroform andwashed with dilute sodium bicarbonate solution and water. The solution,dried over sodium sulfate, is evaporated under vacuum, thus obtaining1.3 g. of17-acetoxy-15β-diethylaminoacetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dioneas a viscous oil.

This oil is taken up in 300 ml. of ether, filtered until clear overcarbon, and combined with ethereal hydrochloric acid until the reactionis acidic as determined by the Congo red indication, thus obtaining 1.6g. of17-acetoxy-15β-diethylaminoacetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dionehydrochloride, melting at about 230° C. under decomposition.

EXAMPLE 10

Analogously to Example 7, 500 mg. of17-acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneis reacted with enanthic anhydride and lauric anhydride, respectively.The following products are obtained:

17-acetoxy-6-chloro-15β-heptanoyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneas an oil, and

17-acetoxy-6-chloro-15β-dodecanoyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dioneas an oil, respectively.

The preceding examples can be repeated with similar success bysubstituting the generically and specifically described reactants and/oroperating conditions of this invention for those used in the precedingexamples.

From the foregoing description, one skilled in the art can easilyascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various usages andconditions.

What is claimed is:
 1. Hydroxylated 1α,2α-methylene steroids of theformula ##STR4## wherein R₁ and R₂ are identical or different and eachis (a) hydrogen; (b) the acyl group of an unsubstituted C₁₋₁₈ alkanecarboxylic or sulfonic acid; an unsubstituted C₆₋₁₀ carbocyclic aromaticcarboxylic or sulfonic acid; an unsubstituted C₇₋₁₈ alkaromaticcarboxylic or sulfonic acid of 1 or 2 aromatic rings; an unsubstitutedC₈₋₁₈ aralkane carboxylic or sulfonic acid of 1 or 2 aromatic rings; ora piperidino, morpholino, pyridine, furan, pyrrolidino or piperazinocarboxylic or sulfonic acid; (c) said acyl groups of (b) substituted byCOOH, lower alkyl, OH, lower alkoxy, oxo, amino or halogen; (d) for saidC₂₋₆ acyl groups of (c) which are substituted by OH, the correspondingacyl groups wherein the OH is esterified by a C₂₋₁₈ alkanoyl or C₂₋₁₈alkyl sulfonyl group; (e) an esterifying group of a hydrohalic,sulphuric or phosphoric acid;and for said 1α,2α-methylene steroids whichare capable of salt formation, the physiologically acceptable saltsthereof. 2.17-Acetoxy-6-chloro-15β-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 3.6-Chloro-15β,17-dihydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione, acompound of claim
 1. 4.17-Acetoxy-15β-adipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 5.17-Acetoxy-15β-adipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,sodium salt, a compound of claim
 1. 6.17-Acetoxy-6-chloro-15β-malonyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 7.17-Acetoxy-6-chloro-15β-malonyloxy-1α,2.alpha.-methylene-4,6-pregnadiene-3,20-dione,sodium salt, a compound of claim 1.8.15β-Adipoyloxy-6-chloro-17-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 9.15β-Adipoyloxy-6-chloro-17-hydroxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,sodium salt, a compound of claim
 1. 10.15β,17-Diadipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 11.15β,17-Diadipoyloxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,disodium salt, a compound of claim
 1. 12.15β,17-Diacetoxy-6-chloro-1.alpha.,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 13. Sodium(17-acetoxy-6-chloro-1α,2α-methylene-3,20-dioxo-4,6-pregnadien-15β-yl)sulfate, a compound of claim
 1. 14.17-Acetoxy-6-chloro-15β-chloroacetoxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 15.15-Acetoxy-15β-diethylaminoacetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 16.17-Acetoxy-15β-diethylaminoacetoxy-6-chloro-1α,2α-methylene-4,6-pregnadiene-3,20-dionehydrochloride, a compound of claim
 1. 17.17-Acetoxy-6-chloro-15β-heptanoyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 18.17-Acetoxy-6-chloro-15β-dodecanoyloxy-1α,2α-methylene-4,6-pregnadiene-3,20-dione,a compound of claim
 1. 19. A compound of claim 1, wherein the acid is aC₁₋₁₈ organic carboxylic or sulfonic acid.
 20. A compound of claim 1,wherein the acid is sulfuric or phosphoric acid.
 21. A compound of claim1, wherein the acid is formic acid, acetic acid, propionic acid, butyricacid, isobutyric acid, valeric acid, isovaleric acid, caproic acid,enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylicacid, trimethylacetic acid, diethylacetic acid, cyclopentylpropionicacid, cyclohexylacetic acid, phenylacetic acid, benzoic acid, glycolicacid, glycolic acid O-undecanoate, mono-, di-, and trichloroacetic acid,oxalic acid, malonic acid, succinic acid, adipic acid, pimelic acid,suberic acid, aminoacetic acid, diethylaminoacetic acid,diethylaminopropionic acid, piperidinoacetic acid, morpholinoaceticacid, nicotinic acid, isonicotinic acid, furan-2-carboxylic acid,methanesulfonic acid, ethanesulfonic acid, isopropanesulfonic acid,butanesulfornic acid, cyclopropanesulfonic acid, cyclopentanesulfonicacid, benzenesulfonic acid, p-toluenesulfonic acid,p-chlorobenzenesulfonic acid, N,N-dimethylainosulfonic acid,N,N-diethylaminosulfonic acid or pyrrolidino-, piperidino-, piperazino-,N-methylpiperazino- or morpholinosulfonic acid.
 22. A compound of claim1, which is an acid addition salt of an aminocarboxylic or sulfonic acidmoiety in the R₁ or R₂ positions; or a basic salt of a dicarboxylic acidmonoester moiety in the R₁ or R₂ positions.
 23. A compound of claim 1,which is a basic salt of a hemisulfate or monophosphoric acid estermoiety in the R₁ or R₂ positions.
 24. A pharmaceutical compositioncomprising an antiandrogenically effective amount of a compound of claim1 and a pharmaceutically acceptable carrier.
 25. A method of treatingandrogenic diseases in mammals which comprises administering to a mammalan antiandrogenically effective amount of a compound of claim
 1. 26. Themethod of claim 25 wherein the disease is acne or seborrhea.
 27. Thepharmaceutical composition of claim 24, wherein the amount ofantiandrogenic compound is 10-100 mg.
 28. The pharmaceutical compositionof claim 24, wherein the amount of antiandrogenic compound is 0.001 to 1wt.%.
 29. The pharmaceutical composition of claim 24, wherein thecarrier is viscous to semi-solid or solid in consistency.